Figure 1a: Experimental plate with DNA plasmid. The plate contains agar with ampicillin and LB media, and a solution of SOC media, BL21(DE3) E.coli bacteria, and pGEM-gbr22 DNA plasmid. The plate sat in a 37-degree Celcius incubator for 16 hours. The red and black marks are from colored markers.
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Figure 1b: Control plate without DNA plasmid. The agar plate is made of LB media and ampicillin with a solution of SOC media and BL21(DE3) E. coli bacteria on top. Plate sat in a 37-degree Celcius incubator for 16 hours. The red and black faded marks are made by colored markers.
Figure 1c: Fun plate with sample taken from a sponge in the lab sink. The agar is a mixture of agar and LB media (no ampicillin). Plate was inside a 37-degree Celsius incubator for 36 hours. The white and red specks are from solid dirt and marker, respectively, on the outside of the dish.
Figure 1d: Experimental plate with DNA plasmid from another group. The agar is a mixture of agar, LB media, and ampicillin. A solution of SOC media, BL21(DE3) E. coli bacteria, and pGEM-gbr22 DNA plasmid are rolled on top. Plate was placed in a 37-degree Celsius incubator for 16 hours. The white specks are bacteria colonies; the red marks are made by a colored marker.
Figure 2: Culture solution with purple protein. This solution contains LB media, ampicillin, BL21(DE3) E. coli bacteria, and pGEM-gbr22 DNA plasmid. This sample was left in the 37-degree Celsius shaking incubator for 22 hours. The purple color inside indicates the presence of proteins in the solution.
Figure 3: Purple protein separated from solution after centrifuge. The pellet is made from a solution of LB media, ampicillin, BL21(DE3) E. coli bacteria, and pGEM-gbr22 DNA plasmid. Image was taken after straining the leftover liquid solution from the conical tube. This pellet weighed 0.70 g.
Figure 4: Elution 1 from purified pGEM-gbr22 purple protein. This solution was filtered through an Econo column with Ni2+ and the addition of 250 mM elution buffer. The purple color indicates the presence of the purple protein in the solution.
Figure 5: Elution 2 from purified pGEM-gbr22 purple protein. This solution has a light purple hue to it, which indicates the small presence of purple protein pGEM-gbr22.
Figure 6: Absorbance vs. Wavelength of Elution 1 at 280 nm (Protein A-280).Trial 1 for the measurement of absorbance of Elution 1 with 280 nm gives a value of 0.23. The blue vertical line on the graph represents where 280 nm is. The black line traces the absorbance at different wavelengths.
Figure 7: Absorbance vs. Wavelength of Elution 1 at 280 nm (Protein A-280). Trial 2 for Elution 1 gives the absorbance value of the solution at 0.245 with the mg/mL of 0.25. The blue vertical line indicates where 280 nm is on the graph. The black line traces the absorbance at different values of wavelength.
Figure 8: Absorbance vs. Wavelength of Elution 2 at 280 nm (Protein A-280). The first trial for Elution 2 has an absorbance value of 0.031. The concentration (mg/mL) is 0.03. The blue vertical line indicates where 280 nm is on the graph. The black line traces the absorbance values at different wavelengths.
Figure 9: Absorbance vs. Wavelength of Elution 2 at 280 nm (Protein A-280).The second trial for Elution 2 has an absorbance value of 0.010 with a mg/mL of 0.01. The blue vertical line indicates where 280 nm is on the graph. The black line traces the absorbance values at different wavelengths.
Figure 10: Absorbance vs. Wavelength of Elution 1 at 574 nm (UV/Vis). The Elution 1 absorbed 0.033 from 574 nm wavelength and 0.023 at 280 nm. The black vertical line represents 574 nm and the blue one represents 280 nm on the graph. The black line traces the absorbance values at different wavelengths (Trial 1). Figure 11: Absorbance vs. Wavelength of Elution 1 at 574 nm (UV/Vis). Trial 2 of measuring the absorbance of Elution 1 at 574 nm has an absorbance value of 0.035. The absorbance at 280 nm is 0.023. The black vertical line represents 574 nm on the graph, while the blue line represents 280 nm. The black line traces the absorbance at different wavelengths. Figure 12: Stained PAGE gel with pGEM-gbr22 purple protein from 8 different samples. The ladder serves as a comparison for the bands on the other columns. The length of the purple protein is represented by the fourth blue line underneath the red line on the ladder. Elution 1 and 2 from the first and second column belong to a different group than Elution 1 and 2 on the 3rd and 4th column.
Figure 13:Example of Thermo Scientific Spectra Multicolored Broad Range Protein Ladder that can be used as a reference for weight comparison of other proteins.
Introduction:
Materials & Methods:
Results:
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Figure 1b: Control plate without DNA plasmid. The agar plate is made of LB media and ampicillin with a solution of SOC media and BL21(DE3) E. coli bacteria on top. Plate sat in a 37-degree Celcius incubator for 16 hours. The red and black faded marks are made by colored markers.Figure 2: Culture solution with purple protein. This solution contains LB media, ampicillin, BL21(DE3) E. coli bacteria, and pGEM-gbr22 DNA plasmid. This sample was left in the 37-degree Celsius shaking incubator for 22 hours. The purple color inside indicates the presence of proteins in the solution.
Figure 3: Purple protein separated from solution after centrifuge. The pellet is made from a solution of LB media, ampicillin, BL21(DE3) E. coli bacteria, and pGEM-gbr22 DNA plasmid. Image was taken after straining the leftover liquid solution from the conical tube. This pellet weighed 0.70 g.
Trial 2 for Elution 1 gives the absorbance value of the solution at 0.245 with the mg/mL of 0.25. The blue vertical line indicates where 280 nm is on the graph. The black line traces the absorbance at different values of wavelength.
The first trial for Elution 2 has an absorbance value of 0.031. The concentration (mg/mL) is 0.03. The blue vertical line indicates where 280 nm is on the graph. The black line traces the absorbance values at different wavelengths.
The Elution 1 absorbed 0.033 from 574 nm wavelength and 0.023 at 280 nm. The black vertical line represents 574 nm and the blue one represents 280 nm on the graph. The black line traces the absorbance values at different wavelengths (Trial 1).
Figure 11: Absorbance vs. Wavelength of Elution 1 at 574 nm (UV/Vis).
Trial 2 of measuring the absorbance of Elution 1 at 574 nm has an absorbance value of 0.035. The absorbance at 280 nm is 0.023. The black vertical line represents 574 nm on the graph, while the blue line represents 280 nm. The black line traces the absorbance at different wavelengths.
Figure 12: Stained PAGE gel with pGEM-gbr22 purple protein from 8 different samples.
The ladder serves as a comparison for the bands on the other columns. The length of the purple protein is represented by the fourth blue line underneath the red line on the ladder. Elution 1 and 2 from the first and second column belong to a different group than Elution 1 and 2 on the 3rd and 4th column.
Figure 13: Example of Thermo Scientific Spectra Multicolored Broad Range Protein Ladder that can be used as a reference for weight comparison of other proteins.
Discussion:
Conclusions:
References: